PURPOSE:Transient receptor potential vanilloid 4 (TRPV4) channels are mechanosensitive ion channels implicated in Ca2+ signalling across various tissues, yet their functional role in human lens epithelial cells remains poorly understood. We aimed to investigate the contribution of TRPV4 to mechanically induced Ca2+ signalling in human anterior lens epithelium. METHODS:Human anterior lens epithelial preparations were obtained from postoperative lens capsules, including both cataractous and non-cataractous samples. Using multicellular Ca2+ imaging, we analysed the spatiotemporal properties of intercellular Ca2+ waves following localised mechanical stimulation under control conditions and after pharmacological inhibition of TRPV4 with HC-067047. RESULTS:Mechanical stimulation triggered radially propagating Ca2+ waves characterised by distance-dependent attenuation of amplitude and kinetics. TRPV4 inhibition significantly reduced wave propagation velocity and enhanced spatial signal decay, indicating a key role in maintaining efficient intercellular Ca2+ signal propagation. These effects were most pronounced in non-cataractous epithelium, while in cataractous lenses, they exhibited stage-dependent differences, suggesting remodelling of Ca2+ signalling pathways during disease progression. The observed intercellular Ca2+ wave propagation is consistent with gap junctional and paracrine signalling, while TRPV4 activity modulates its efficiency by shaping the initiating Ca2+ response. CONCLUSION:TRPV4 channels contribute to both the efficiency and spatial extent of Ca2+ signalling in the lens epithelium. These findings identify TRPV4 as an important regulator of mechanically induced intercellular communication and a potential target for modulating lens function in health and disease.

Purpose: To determine the synchrotron radiation‐based FTIR microspectroscopy spectrum of lens capsules (LCs) from cataract surgery and that of idiopathic epiretinal membranes (iERMs) towards finding a possible causal relationship between lens fragments falling onto the posterior segment during phacoemulsification procedure and development of iERMs.Methods: LCs from cataract surgery and iERMs were collected from 5 patients. FTIR measurements were performed at the MIRAS beamline of the ALBA Synchrotron, Barcelona, Spain, and the complete bio‐macromolecular spectral information at cellular level (proteins, lipids, and nucleic acids) in the lens epithelium of LCs from cataract surgery and iERMs were evaluated and compared.Results: The FTIR spectra revealed several bands corresponding to the vibration of various groups associated with proteins, lipids, and nucleic acids, demonstrating the complex biochemical composition of the LCs and iERMs. Overall, the spectra of the LCs and iERMs were very similar, showing slight differences in the spectral region of the proteins, particularly in the Amide I, then in the carbonyl groups, and phosphate groups of DNA. Additionally, the lipid region showed differences in the spectral ratio between the LCs and iERMs, characterized by CH2 and CH3 bands.Conclusions: The FTIR spectra of lens epithelial cells on LCs and cells within the iERMs appeared similar, with slight differences in some spectral regions. It is possible that the lens fragments falling onto the retina during cataract surgery could contribute to the pathogenesis of iERMs.

Purpose:Effective intercellular communication among lens epithelial cells (LECs) is essential for lens homeostasis, and its disruption has been implicated in cataract formation. This study investigates the mechanisms of calcium (Ca²⁺) wave propagation in the human lens epithelium, focusing on the respective roles of gap-junctional coupling and ATP-mediated paracrine signaling. Methods:We performed multicellular Ca²⁺ imaging on human postoperative anterior lens capsule preparations obtained from cataractous lenses during cataract surgery which retained intact monolayers containing viable LECs. Mechanically induced Ca²⁺ waves were recorded, and the contribution of specific signaling pathways was evaluated by pharmacological intervention using apyrase (an ATP-hydrolyzing enzyme) and carbenoxolone (CBX; a gap-junctional blocker). To interpret the experimental results, we developed a biophysically detailed computational model of the LEC monolayer, incorporating intracellular Ca²⁺ dynamics, gap-junctional IP₃/Ca²⁺ diffusion, and extracellular ATP signaling. Results:Apyrase moderately reduced the spatial extent, amplitude, and duration of Ca²⁺ waves without affecting propagation speed. In contrast, CBX significantly suppressed wave transmission, limiting activation to cells directly adjacent to the stimulation site. Simulations reproduced key experimental features and indicated that neither pure gap-junctional nor purely paracrine signaling mechanisms alone could explain the observed dynamics. Instead, a hybrid mechanism combining gap-junctional communication and partially regenerative ATP release was required. Conclusions:Our results highlight the cooperative roles of gap-junctional and ATP-based paracrine signaling in mediating mechanically induced Ca²⁺ wave propagation in the human lens epithelium. This dual-pathway mechanism may be critical for coordinated cellular responses that support physiological processes such as ion homeostasis and transparency maintenance in the human lens.

Human primary lens epithelial cultures serve as an in vitro model for posterior capsular opacification (PCO) formation. PCO occurs when residual lens epithelial cells (LECs) migrate and proliferate after cataract surgery, differentiating into fibroblastic and lens fiber-like cells. This study aims to show and compare the bio-macromolecular profiles of primary LEC cultures and postoperative lens epithelia LECs on basal laminas (bls), while also analyzing bls and cultured LECs separately. Using synchrotron radiation-based Fourier transform infrared (SR-FTIR) (Bruker, Karlsruhe, Germany) microspectroscopy at the Spanish synchrotron light source ALBA, we observed that the SR-FTIR measurements were predominantly influenced by the strong collagen absorbance of the bls. Cultured LECs on bls showed a higher collagen contribution, indicated by higher vas CH3, CH2 and CH3 wagging and deformation, and the C–N stretching of collagen. In contrast, postoperative LECs on bls showed a higher cell contribution, indicated by the vsym CH2 peak and the ratio between vas CH2 and vas CH3 peaks. The primary difference revealed using SR-FTIR is the greater LEC contribution in spectra recorded from postoperative lens epithelia compared to cultured LECs on bls. IR spectra for bl, cultured LECs and postoperative lens epithelia could be valuable for future research.

Aims/Purpose: To compare the bio‐macromolecular composition and the differences between the human cultured and post‐operative lens epithelial cells (LECs) on lens capsules (LCs) for a better understanding of posterior capsular opacification (PCO)—the most common complication of cataract surgery, and lens regeneration. Methods: The explants of the anterior portion of the LC containing the LECs, obtained from cataract surgery and cultivated under adherent conditions, as well as the post‐operative LC with LECs, were analysed by using synchrotron radiation‐based Fourier transform infrared (SR‐FTIR) microspectroscopy, a vibrational spectroscopic technique that allows monitoring of the entire biochemical status of the biological processes. The SR‐FTIR microspectroscopy setup installed on the beamline MIRAS at the Spanish synchrotron light source ALBA was used, where measurements were set to achieve single‐cell resolution, with high spectral stability and high photon flux. Results: We found that the differences exist between the composites of cultured and post‐operative LECs on LCs on the level of all chemical constituents: proteins, lipids, nucleic acids and carbohydrates as well as oxidative stress. The strongest differences are found in protein secondary structure contribution where the composite of primary LECs cultures on LC have more α‐helix (1652 cm −1 ) and post‐operative LC have more β‐sheet (1624 cm −1 and 1697 cm −1 ) secondary structures. The oxidative stress is more expressed in the composite of primary LECs cultures on LC and relatively less in the composite of post‐operative lens epithelium LECs on LC. Conclusions: Our results obtained by SR‐FTIR increase the knowledge about the total proteins, lipids, and nucleic acids in human primary cultures LECs while offering also evidence that SR‐FTIR is sensitive to the pathologic processes of LECs transdifferentiation. We showed that the composite of primary cultures LECs on LC have a distinct bio‐macromolecular composition compared to the composite of post‐operative LECs on LC, which gives additional information for a better understanding of PCO and lens regeneration.

Synchrotron radiation-based Fourier Transform Infrared (SR-FTIR) microspectroscopy is a non-destructive and chemically sensitive technique for the rapid detection of changes in the different components of the cell's biomacromolecular profile. Reactive oxygen species and oxidative stress may cause damage to the DNA, RNA, and proteins in the retinal pigment epithelium (RPE), which can further lead to age-related macular degeneration (AMD) and visual loss in the elderly. In this study, human primary RPEs (hRPEs) were used to study AMD pathogenesis by using an established in vitro cellular model of the disease. Autophagy-a mechanism of intracellular degradation, which is altered during AMD, was studied in the hRPEs by using the autophagy inducer rapamycin and treated with the autophagy inhibitor bafilomycin A1. In addition, oxidative stress was induced by the hydrogen peroxide (H2O2) treatment of hRPEs. By using SR-FTIR microspectroscopy and multivariate analyses, the changes in the phosphate groups of nucleic acids, Amide I and II of the proteins, the carbonyl groups, and the lipid status in the hRPEs showed a significantly different pattern under oxidative stress/autophagy induction and inhibition. This biomolecular fingerprint can be evaluated in future drug discovery studies affecting autophagy and oxidative stress in AMD.

Purpose: To investigate the characteristics of untreated and treated human retinal pigmented epithelial cells (hRPEs) as an ex vivo model for age-related macular degeneration (AMD) using high resolution synchrotron radiation-based Fourier Transform Infrared (FTIR) microspectroscopy and multivariate analysis. Methods: hRPEs obtained from cadavers were treated by autophagy -inducer rapamycin (Rap), and -inhibitor bafilomycin A1 (Baf), as well as hydrogen peroxide (H 2 O 2 ) to induce oxidative stress, in an ex vivo model for studying AMD. FTIR measurements were performed at the MIRAS beamline of the ALBA Synchrotron, Barcelona, Spain. Results: The FTIR spectra consisted of several bands arising from the vibration of different groups belonging to proteins, lipids and nucleic acids, indicating the rich biochemical composition of the hRPEs. In the protein region, the amide I band of the treated groups (Rap, Baf, H 2 O 2 ) was different from the control untreated group, indicating an overall protein disordering under such treatments. In the DNA region, two spectral areas appeared to be modified upon treatment, indicating different modifications of the DNA conformational changes or rearrangements attributed to the distortions of the DNA double helix in the presence of Rap, Baf or H 2 O 2 . The same treatments could also induce several modifications in the lipid spectral region, which can affect a wide range of biological processes. The H 2 O 2 treated group showed higher prevalence for the lipid peroxidation, a process generated by the effect of several reactive oxygen species. Conclusions: The present study shows that high resolution FTIR can be used to evaluate the complete bio−/macro-molecular spectra of hRPEs such as proteins, lipids and nucleic acids, which can be used as an ex vivo model for AMD, thus giving possibilities for developing and testing new treatment modalities.

Pathological tissue on the surface of the retina that can be of different etiology and pathogenesis can cause changes in the retina that have a direct consequence on vision. Tissues of different etiology and pathogenesis have different morphological structures and also different macromolecule compositions usually characteristic of specific diseases. In this study, we evaluated and compared biochemical differences among samples of three different types of epiretinal proliferations: idiopathic epiretinal membrane (ERMi), membranes in proliferative vitreoretinopathy (PVRm), and proliferative diabetic retinopathy (PDRm). The membranes were analyzed by using synchrotron radiation-based Fourier transform infrared micro-spectroscopy (SR-FTIR). We used the SR-FTIR micro-spectroscopy setup, where measurements were set to achieve a high resolution that was capable of showing clear biochemical spectra in biological tissue. We were able to identify differences between PVRm, PDRm, and ERMi in protein and lipid structure; collagen content and collagen maturity; differences in proteoglycan presence; protein phosphorylation; and DNA expression. Collagen showed the strongest expression in PDRm, lower expression in ERMi, and very low expression in PVRm. We also demonstrated the presence of silicone oil (SO) or polydimethylsiloxane in the structure of PVRm after SO endotamponade. This finding suggests that SO, in addition to its many benefits as an important tool in vitreoretinal surgery, could be involved in PVRm formation.

Purpose: To assess epiretinal proliferations bio-macromolecules and to provide their molecular fingerprint for better understanding of epiretinal proliferations, we used synchrotron radiation-based Fourier transform infrared (SR-FTIR) micro-spectroscopy. Methods: The membranes were collected from routine pars plana vitrectomy. The biochemical differences between idiopathic nonvascular epiretinal membranes (ERMi), membranes in proliferative vitreoretinopathy (PVRm) and neovascular membranes in proliferative diabetic retinopathy (PDRm) were evaluated and compared by using SR-FTIR micro-spectroscopy at the MIRAS beamline of the ALBA Synchrotron, Barcelona, Spain and multivariate analysis. Results: PVRm differs the most from ERMi and PDRm in all spectral regions. We demonstrated the presence of silicone oil (SO) or polydimethylsiloxane (PDMS) in the structure of PVRm membrane after SO endotamponade suggesting that SO, in addition to many benefits as an important tool in vitreoretinal surgery, can also contribute in PVRm formation. We also showed the differences between PVRm, PDRm and ERMi in protein and lipid structure, collagen content and maturity. Conclusions: Our results obtained by SR-FTIR increase the knowledge about the total proteins, lipids and nucleic acids in different epiretinal proliferations while offering also the evidence that SR-FTIR is sensitive to the pathologic processes of epiretinal proliferations. We emphasize the different macro-molecular composition of three different types of epiretinal proliferation stemming from different components and metabolic processes taking place as well as the caution needed when using SO in vitreoretinal surgery.

Oxidative stress is caused by an imbalance between reactive oxygen species (ROS) generation and the capacity of antioxidant ROS scavenging systems and plays an essential role in the pathogenesis of many diseases. It is connected with cell damage, such as lipid peroxidation of membranes. One important source of oxidative stress is UV radiation, which can come from the natural environment or artificial sources like welding. While sources of artificial UV radiation emit specific wavelengths depending on the application, occupational exposure to natural UV radiation has a continuous spectrum from 290 nm to 400 nm. Oxidative stress can be measured by synchrotron radiation-based Fourier Transform Infrared (SR-FTIR) microspectroscopy. Oxidative effect of UV can be studied on human postoperative tissue. Here we show an in vitro study of the effect of UV C on the oxidative stress in human eye postoperative tissue.

Ca2+ homeostasis and signaling disturbances are associated with lens pathophysiology and are involved in cataract formation. Here, we explored the spatiotemporal changes in Ca2+ signaling in lens epithelial cells (LECs) upon local mechanical stimulation, to better understand the LECs’ intercellular communication and its association with cataractogenesis. We were interested in if the progression of the cataract affects the Ca2+ signaling and if modifications of the Ca2+ homeostasis in LECs are associated with different cataract types. Experiments were done on the human postoperative anterior lens capsule (LC) preparations consisting of the monolayer of LECs on the basement membrane. Our findings revealed that the Ca2+ signal spreads radially from the stimulation point and that the amplitude of Ca2+ transients decreases with increasing distance. It is noteworthy that a comparison of signaling characteristics with respect to the degree of cataract progression revealed that, in LCs from more developed cataracts, the Ca2+ wave propagates faster and the amplitudes of Ca2+ signals are lower, while their durations are longer. No differences were identified when comparing LCs with regard to the cataract type. Moreover, experiments with Apyrase have revealed that the Ca2+ signals are not affected by ATP-dependent paracrine communication. Our results indicated that cataract progression is associated with modifications in Ca2+ signaling in LECs, suggesting the functional importance of altered Ca2+ signaling of LECs in cataractogenesis.

Ultraviolet (UV) irradiation is an important risk factor in cataractogenesis. Lens epithelial cells (LECs), which are a highly metabolically active part of the lens, play an important role in UV-induced cataractogenesis. The purpose of this study was to characterize cell compounds such as nucleic acids, proteins, and lipids in human UV C-irradiated anterior lens capsules (LCs) with LECs, as well as to compare them with the control, non-irradiated LCs of patients without cataract, by using synchrotron radiation-based Fourier transform infrared (SR-FTIR) micro-spectroscopy. In order to understand the effect of the UV C on the LC bio-macromolecules in a context of cataractogenesis, we used the SR-FTIR micro-spectroscopy setup installed on the beamline MIRAS at the Spanish synchrotron light source ALBA, where measurements were set to achieve a single-cell resolution with high spectral stability and high photon flux. UV C irradiation of LCs resulted in a significant effect on protein conformation with protein formation of intramolecular parallel β-sheet structure, lower phosphate and carboxyl bands in fatty acids and amino acids, and oxidative stress markers with significant increase of lipid peroxidation and diminishment of the asymmetric CH3 band.

The purpose of this work is to examine the structure of the anterior lens epithelial cells (aLECs) of presenile idiopathic cortical cataract to investigate the possible structural reasons for its development. The anterior lens capsules (aLCs: basement membrane and associated lens epithelial cells) were obtained from routine uneventful cataract surgery of 5 presenile cataract patients (16 and 41 years old women and 29, 39, and 45 years old men). None of the patients had family history of cataract, medication, or trauma and they were otherwise healthy. In addition, the patients did not have any other abnormal features in the ocular status except cataract. The aLCs were prepared for scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The most prominent abnormal features observed by SEM for all 5 studied presenile cataract patients were the changes of the aLECs structure with the dents, the selective concavity of some LECs, at their apical side centrally toward the nucleus. In addition, TEM showed the thinning of the lens epithelium with the segmentally concave cells and the compressed and elongated nuclei. Abnormal and distinguishable structural features were observed in the anterior lens epithelium aLECs in all 5 patients with presenile cataract. Disturbed structure of aLECs, regularly present in presenile cataract type is shown that might be associated with water accumulation in the presenile idiopathic cortical cataract lens.

Cataract is the leading cause of blindness worldwide but the mechanisms involved in the process of cataractogenesis are not yet fully understood. Two most prevalent types of age-related cataracts are nuclear (N) and cortical (C) cataracts. A common environmental factor in most age-related cataracts is believed to be oxidative stress. The lens epithelium, the first physical and biological barrier in the lens, is build from lens epithelial cells (LECs). LECs are important for the maintenance of lens transparency as they control energy production, antioxidative mechanisms and biochemical transport for the whole lens. The purpose of this study is to characterize compounds in LECs originated from N and C cataracts, by using the synchrotron radiation-based Fourier Transform Infrared (SR-FTIR) microspectroscopy, in order to understand the functional importance of their different bio-macromolecules in cataractogenesis. We used the SR-FTIR microspectroscopy setup installed on the beamline MIRAS at the Spanish synchrotron light source ALBA, where measurements were set to achieve single cell resolution, with high spectral stability and high photon flux. The results showed that protein aggregation in form of fibrils was notably pronounced in LECs of N cataracts, while oxidative stress and the lipids peroxidation were more pronounced in LECs of C cataracts.

The mechanisms involved in the process of cataract formation are not yet fully understood. Lens epithelial cells (LECs) are important for the maintenance of lens transparency as they control energy production, antioxidative mechanisms and biochemical transport for the whole lens. The purpose of this study is to characterize LECs compounds of two different cataract types, nuclear (N) and cortical (C), by using the high resolution synchrotron radiation-based Fourier Transform Infrared (SR-FTIR) microspectroscopy. The anterior lens capsule (basement membrane and associated LECs) were obtained from cataract surgery and prepared by desiccation for FTIR. Measurements were set to achieve single cell resolution. Data obtained by SR-FTIR have been analysed by comparing all spectra of N type and C type LECs. The average spectra of each group have been calculated and principal component analysis (PCA) has been performed. The first component of the PCA in Amide I area of spectra showed the most prominent contribution at 1625 cm -1 which is associated with beta sheet organisation in protein aggregates in N cataract type. In the C type we found proteins organized more in the turn and loops conformation (~1670 cm -1 ). The markers of oxidative stress: ratios of the asymmetric vibration CH 2 and CH 3 , as well as C=O/lipids showed an increase in C cataract type LECs that pointed out to an increase in lipid peroxidation in this type of LECs. The region corresponds to the P=O asymmetric bands indicate possible difference in protein phosphorylation in C and N types LECs. Here we evaluated the proteins conformation changes, as well as lipids and phosphates bands in LECs of two different cataract types. The oxidative stress and the lipids peroxidation were more pronounced in C type, while protein aggregation is associated with N type. These spectral changes can be assigned to specific biochemical processes occur in two different cataract types LECs.

Posterior capsule opacifcation–PCO is the most common complication after cataract surgery. Proliferation and migration of lens epithelial cells that remain in the capsular bag following cataract surgery can lead to the development of PCO, which is the main cause of deterioration of visual function. PCO shows the classic features of fbrosis, including hyperproliferation, migration, deposition of matrix and its shrinkage and transdifferentiation into myofbroblast. Astonishingly, the results of recent research show the importance of lens epithelium for lens regeneration following congenital cataract surgery. New minimally invasive cataract surgery removes only 1–1.5 mm of lens epithelium more laterally, so the major part of the epithelium remains in the capsular bag. Conceptually, the new method differs from the current practice, since it preserves the endogenous epithelial cells of the lens and achieves functional lens regeneration in rabbits and monkeys as well as in human infants with congenital cataracts. Pluripotency of lens epithelial cells and their stem cell nature are crucial for lens regeneration.Ex vivo cultures of the lens capsule explants can be used for testing the pharmacological agents for stimulating and inhibiting the growth of lens epithelial cells. Functionality of the cells and responses to pharmacological agents can be studied by analyzing the intra- and extra-cellular calcium (Ca2+) signaling. Stimulating the growth of lens epithelial cells is important in lens regeneration while inhibiting the growth of lens epithelial cells is important in preventing the development of PCO. In the article I described the methods for the analysis of lens epithelial cells after cataract surgeries, which are carried out in the laboratory of the Eye Hospital, University Medical Centre Ljubljana.

Posterior capsule opacifcation–PCO is the most common complication after cataract surgery. Proliferation and migration of lens epithelial cells that remain in the capsular bag following cataract surgery can lead to the development of PCO, which is the main cause of deterioration of visual function. PCO shows the classic features of fbrosis, including hyperproliferation, migration, deposition of matrix and its shrinkage and transdifferentiation into myofbroblast. Astonishingly, the results of recent research show the importance of lens epithelium for lens regeneration following congenital cataract surgery. New minimally invasive cataract surgery removes only 1–1.5 mm of lens epithelium more laterally, so the major part of the epithelium remains in the capsular bag. Conceptually, the new method differs from the current practice, since it preserves the endogenous epithelial cells of the lens and achieves functional lens regeneration in rabbits and monkeys as well as in human infants with congenital cataracts. Pluripotency of lens epithelial cells and their stem cell nature are crucial for lens regeneration.Ex vivo cultures of the lens capsule explants can be used for testing the pharmacological agents for stimulating and inhibiting the growth of lens epithelial cells. Functionality of the cells and responses to pharmacological agents can be studied by analyzing the intra- and extra-cellular calcium (Ca2+) signaling. Stimulating the growth of lens epithelial cells is important in lens regeneration while inhibiting the growth of lens epithelial cells is important in preventing the development of PCO. In the article I described the methods for the analysis of lens epithelial cells after cataract surgeries, which are carried out in the laboratory of the Eye Hospital, University Medical Centre Ljubljana.

PURPOSE:Our purpose was to study the structure of the lens epithelial cells (LECs) of intumescent white cataracts (IC) in comparison with nuclear cataracts (NC) in order to investigate possible structural reasons for development of IC.METHODS:The anterior lens capsule (aLC: basement membrane and associated LECs) were obtained from cataract surgery and prepared for scanning electron microscopy (SEM) and transmission electron microscopy (TEM).RESULTS:We observed by SEM that in IC, LEC swelling was pronounced with the clefts surrounding the groups of LECs. Another structural feature was spherical formations, that were observed on the apical side of LEC's, towards the fibre cell layer, both by SEM and TEM. Development of these structures, bulging out from the apical cell membrane of the LEC's and disrupting it, could be followed in steps towards the sphere formation. The degeneration of the lens epithelium and the structures of the aLC in IC similar to Morgagnian globules were also observed. None of these structural changes were observed in NC.CONCLUSIONS:We show by SEM and TEM that, in IC, LECs have pronounced structural features not observed in NC. This supports the hypothesis that the disturbed structure of LECs plays a role in water accumulation in the IC lens. We also suggest that, in IC, LECs produce bulging spheres that represent unique structures of degenerated material, extruded from the LEC.

Inducing selective or targeted cell apoptosis without affecting large number of neighbouring cells remains a challenge. A plausible method for treatment of posterior capsular opacification (PCO) due to remaining lens epithelial cells (LECs) by reactive chemistry induced by localized single electrode microplasma discharge at top of a needle-like glass electrode with spot size ~3 μm is hereby presented. The focused and highly-localized atmospheric pressure microplasma jet with electrode discharge could induce a dose-dependent apoptosis in selected and targeted individual LECs, which could be confirmed by real-time monitoring of the morphological and structural changes at cellular level. Direct cell treatment with microplasma inside the medium appeared more effective in inducing apoptosis (caspase 8 positivity and DNA fragmentation) at a highly targeted cell level compared to treatment on top of the medium (indirect treatment). Our results show that single cell specific micropipette plasma can be used to selectively induce demise in LECs which remain in the capsular bag after cataract surgery and thus prevent their migration (CXCR4 positivity) to the posterior lens capsule and PCO formation.

PURPOSE:In retinitis pigmentosa (RP) patients, relatively minor lens opacity in central part of posterior pole of the lens may cause disproportionate functional symptoms requiring cataract operation. To investigate the possible structural reasons for this opacity development, we studied the structure of the lens epithelium of patients with RP.METHODS:The anterior lens capsule (aLC: basement membrane and associated lens epithelial cells, LECs) was obtained from cataract surgery and prepared for scanning and transmission electron microscopy (SEM and TEM).RESULTS:Both SEM and TEM show a number of abnormal features in the anterior lens epithelium of cataract patients with RP. The abnormalities appear mainly as holes, thinning and degradation of the epithelium, with the dimensions from <1 μm to more than 50 μm. Other types of holes in size up to 20 μm were seen that may be formed by gradual stretching of the lens epithelium. Another type of abnormalities was cracks that were seen between adjacent LECs, with dimensions 0.1-2 μm × up to 10 μm.CONCLUSIONS:Abnormal structural features were observed in the anterior lens epithelium that may cause water influx into the lens. This may lead to clouding along the water clefts leading towards the posterior pole in the RP cataractous lens. We suggest that the lens epithelium has a role in the development of the cataract in patients with RP.